A peer-reviewed open-access journal ZooKeys 496:Description 85–103 (2015) of a new species of Wormaldia from Sardinia and a new Drusus species... 85 doi: 10.3897/zookeys.496.9169 RESEARCH ARTICLE http://zookeys.pensoft.net Launched to accelerate biodiversity research Description of a new species of Wormaldia from Sardinia and a new Drusus species from the Western Balkans (Trichoptera, Philopotamidae, Limnephilidae) Simon Vitecek1, Ana Previšić2, Mladen Kučinić2, Miklós Bálint3, Lujza Keresztes4, Johann Waringer1, Steffen U. Pauls3, Hans Malicky5, Wolfram Graf6 1 Department of Limnology and Oceanography, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria 2 Department of Biology, Faculty of Science, University of Zagreb, Rooseveltov trg 6, HR-10000 Zagreb, Croatia 3 Senckenberg Biodiversity and Climate Research Centre (BiK-F), Senckenberganlage 25, D-60325 Frankfurt a.M., Germany 4 Hungarian Department of Biology and Ecology, Babeş-Bolyai University, Clinicilor 5–7, 400006 Cluj-Napoca, Romania 5 Sonnengasse 13, Lunz am See, Austria 6 Institute of Hydrobiology and Aquatic Ecology Management, University of Natural Resources and Life Sciences, Max Emanuel-Strasse 17, A-1180 Vienna, Austria Corresponding author: Simon Vitecek ([email protected]) Academic editor: R. Holzenthal | Received 22 December 2014 | Accepted 26 March 2015 | Published 16 April 2015 http://zoobank.org/DE6EE3CA-BEFB-4D20-A411-51743759BCB8 Citation: Vitecek S, Previšić A, Kučinić M, Bálint M, Keresztes L, Waringer J, Pauls SU, Malicky H, Graf W (2015) Description of a new species of Wormaldia from Sardinia and a new Drusus species from the Western Balkans (Trichoptera, Philopotamidae, Limnephilidae). ZooKeys 496: 85–103. doi: 10.3897/zookeys.496.9169 Abstract New species are described in the genera Wormaldia (Trichoptera, Philopotamidae) and Drusus (Trichoptera, Limnephilidae, Drusinae). Additionally, the larva of the new species Drusus crenophylax sp. n. is described, and a key provided to larval Drusus species of the bosnicus-group, in which the new species belongs. Obser- vations on the threats to regional freshwater biodiversity and caddisfly endemism are discussed. The new species Wormaldia sarda sp. n. is an endemic of the Tyrrhenian island of Sardinia and differs most conspicuously from its congeners in the shape of segment X, which is trilobate in lateral view. The new species Drusus crenophylax sp. n. is a micro-endemic of the Western Balkans, and increases the end- emism rate of Balkan Drusinae to 79% of 39 species. Compared to other Western Balkan Drusus, males of the new species are morphologically most similar to D. discophorus Radovanovic and D. vernonensis Malicky, but differ in the shape of superior and intermediate appendages. The females ofD. crenophylax sp. n. are most similar to those of D. vernonensis, but differ distinctly in the outline of segment X. Larvae of D. crenophylax sp. n. exhibit toothless mandibles, indicating a scraping-grazing feeding ecology. Keywords Caddisfly, Europe, larval key, taxonomy, conservation, Mediterranean, hydropower Copyright Simon Vitecek et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 86 Simon Vitecek et al. / ZooKeys 496: 85–103 (2015) Introduction The Mediterranean area is a flora and fauna biodiversity hot-spot. The Tyrrhenian- is lands and the Balkans, in particular, are noteworthy for their high number of plant en- demics (Médail and Quezél 1997, 1999; Nikolić et al. 2008; Fenu et al. 2010; Bacchet- ta et al. 2012), and mammal and invertebrate endemics (Holdhaus 1924, Vigne 1992, Muccedda et al. 2002, Griffiths et al. 2004, Grill et al. 2007). Freshwater biodiversity has recently become a focus of attention throughout Europe, including the Mediter- ranean region with the Western Balkans and Sardinia (e.g., di Sabatino 2003, Zakšek et al. 2009, Tierno de Figueroa et al. 2013, Klobučar et al. 2013, Weiss et al. 2014). The genus Wormaldia currently comprises 204 species (Morse 2014) of which 36 species occur in Europe (Malicky 2005, Graf et al. 2008). Most species are widely dis- tributed, but also several apparently highly endemic species have been described (Graf et al. 2008, Martínez-Menéndez and González 2011). Aquatic stages of the genus, with few exceptions, prefer crenal and rhithral sections of alpine to lowland streams, are case- less and behave as passive filter feeders using characteristic nets (Graf et al. 2008). Spe- cies in the genus exhibit characteristic male genitalia, but also comparatively high vari- ability, particularly of the phallic structures (Malicky 2004, Martínez-Menéndez and González 2011, Neu pers. comm.), resulting in the description of several subspecies. The genus Drusus is in the subfamily Drusinae Banks, and comprises 84 species (Mali- cky 2004, 2005; Kučinić et al. 2011a; Oláh 2010, 2011; Oláh and Kovács 2013). Larvae of the group prefer eucrenal to epirhithral sections of cold alpine or montane streams and brooks. Feeding ecology of Drusus larvae is complex, and three different feeding guilds can be distinguished based on the shape of larval mandibles and leg setation: filtering car- nivores, omnivorous shredders, and scraping grazers (Pauls et al. 2008, Graf et al. 2009). Taxonomic richness of Drusinae is particularly high in the Western Balkans, including a high number of micro-endemics (Malicky 2004; Graf et al. 2008; Oláh 2010, 2011; Kučinić et al. 2011a, b; Oláh and Kovács 2013, Previšić et al. 2014a, b). In this paper we describe a new species of Wormaldia and a new grazer Drusus spe- cies, including a key to the hitherto known larval stages of the bosnicus-group, in which Drusus crenophylax sp. n. belongs. Materials and methods Adults were collected using sweep nets and immature stages by handpicking. Collected specimens were stored in 70% and 96% EthOH, for morphological and molecular analyses, respectively. Male and female genitalia were examined after being cleared in either KOH or lactic acid. Nomenclature of male genitalia of Wormaldia McLachlan follows Nielsen (1957, for Wormaldia occipitalis Pictet), nomenclature of male genitalia of Drusus fol- lows Nielsen (1957, for Limnephilus flavicornisFabricius) using the simplifying terms “superior appendages” for the lateral processes of segment X (cerci sensu Snodgrass Description of a new species of Wormaldia from Sardinia and a new Drusus species... 87 Table 1. Detailed list of Drusus specimens used for mtCOI analysis. Abbreviations: M adult male, F female; L larva; U unknown. Species Locality Specimen ID/Stage Accession # Collectors D. crenophylax 44°32.932'N, 17°23.562'E fDsp4501M/M KP793082 Dmitrović, Šukalo D. crenophylax 44°33.003'N, 17°23.580'E fDsp4502L/L KP793083 Dmitrović, Šukalo D. crenophylax 44°33.003'N, 17°23.580'E fDsp4503L/L KP793081 Dmitrović, Šukalo D. crenophylax 44°33.003'N, 17°23.580'E fDsp3401F/F KP793084 Dmitrović, Šukalo D. crenophylax 44°33.003'N, 17°23.580'E fDsp3402F/F KP793085 Dmitrović, Šukalo D. vernonensis 41°0.887'N, 21°10.448'E DdphPEIM1/M KC881524 Kučinić, Graf D. vernonensis 41°0.887'N, 21°10.448'E DdphPEIM2/M KP793087 Kučinić, Graf D. vernonensis 41°0.887'N, 21°10.448'E DdphPEIM3/M KP793086 Kučinić, Graf D. discophorus Macedonia, Jablanica Mts. fDds0110M/M KP793089 Kučinić D. discophorus Macedonia, Jablanica Mts. fDds0112F/F KP793088 Kučinić 1935), and “intermediate appendages” for the sclerite and the anterior process of seg- ment X (paraproct sensu Snodgrass 1935). Nomenclature of larval morphological fea- tures follows Wiggins (1998) and Waringer and Graf (2011), nomenclature of primary setae and setal areas follows Wiggins (1998). Illustrations were prepared according to Thomson and Holzenthal (2010) in which pencil drawings made with a camera lucida are digitized, edited and inked in Adobe Illustrator (v. 16.0.4, Adobe Systems Inc.). Molecular genetic sequence data were used to support larval association and assess relationships to previously described Drusus species. DNA extraction and amplification of a 541-bp-long fragment of the mtCOI gene using standard primers (forward primer: Jerry, Simon et al. 1994, reverse primer: S20, Pauls et al. 2006) was performed as outlined by Pauls et al. (2008) and Previšić et al. (2009b). Sequences were edited manually using Geneious version R7 (http://www.geneious.com, Kearse et al. 2012) and aligned using MAFFT (Katoh and Standley 2013). Sequences were deposited in GenBank under Acces- sion nos: KC881524, KP793081–KP793089 (Table 1). Inter- and intraspecific genetic distances (uncorrected p-distances) were calculated in Mega 4.0.1 (Tamura et al. 2007). Taxonomy Wormaldia sarda Graf & Malicky, sp. n. http://zoobank.org/F02C5CF5-9043-463F-809B-FCD5D2B8FBD2 Material examined. Holotype. 1 male pupa, holotype: Sardinia, Gola di Gorruppo; 40°11.122'N, 9°30.104'E; 350 m a.s.l.; 28.03.2001; leg. Monika Hess, Ulrich Heckes; currently in coll. W. Graf, will deposited in the Biologiezentrum des Oberösterreichis- chen Landesmuseums, Linz, Austria. Type locality. Italy, Sardinia. Diagnosis. Morphology of the male terminalia suggests placement of the new species in Wormaldia. The species is unique in the European Trichoptera fauna, and easily differentiated from all otherWormaldia species by the combination of the fol- 88 Simon Vitecek et al. / ZooKeys 496: 85–103 (2015) lowing characters: (1) presence of median subtriangular